{"doi":"10.1093/nar/gkac589","title":"Flipped over U: structural basis for dsRNA cleavage by the SARS-CoV-2 endoribonuclease","abstract":"Coronaviruses generate double-stranded (ds) RNA intermediates during viral replication that can activate host immune sensors. To evade activation of the host pattern recognition receptor MDA5, coronaviruses employ Nsp15, which is a uridine-specific endoribonuclease. Nsp15 is proposed to associate with the coronavirus replication-transcription complex within double-membrane vesicles to cleave these dsRNA intermediates. How Nsp15 recognizes and processes dsRNA is poorly understood because previous structural studies of Nsp15 have been limited to small single-stranded (ss) RNA substrates. Here we present cryo-EM structures of SARS-CoV-2 Nsp15 bound to a 52nt dsRNA. We observed that the Nsp15 hexamer forms a platform for engaging dsRNA across multiple protomers. The structures, along with site-directed mutagenesis and RNA cleavage assays revealed critical insight into dsRNA recognition and processing. To process dsRNA Nsp15 utilizes a base-flipping mechanism to properly orient the uridine within the active site for cleavage. Our findings show that Nsp15 is a distinctive endoribonuclease that can cleave both ss- and dsRNA effectively.","journal":"Nucleic Acids Research","year":2022,"id":238697,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":53,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9469,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":644957,"name":"Isha M. Wilson","orcid":"0000-0001-5441-7336","position":1,"is_corresponding":false},{"id":398021,"name":"J.M. Krahn","orcid":"0000-0002-4640-0267","position":2,"is_corresponding":false},{"id":806985,"name":"Kevin John Butay","orcid":"0000-0001-8895-5052","position":3,"is_corresponding":false},{"id":559409,"name":"Lucas Dillard","orcid":"0000-0003-4325-8055","position":4,"is_corresponding":false},{"id":227365,"name":"Mario J. Borgnia","orcid":"0000-0001-9159-1413","position":5,"is_corresponding":false},{"id":336384,"name":"Robin E. Stanley","orcid":"0000-0002-2106-3102","position":6,"is_corresponding":false},{"id":336383,"name":"Meredith N. Frazier","orcid":"0000-0003-3205-8832","position":0,"is_corresponding":true}],"reference_count":53,"raw_metadata":null,"created_at":"2026-07-19T00:22:31.741535Z","pmid":"35801916","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}